FTO在睾丸组织中的表达定位及潜在调控基因的筛选研究  

The localization and screening of potential regulatory genes of FTO in testicular tissue

作　　者：沈千怡 范俊杰 黄鑫 蒋莹 何科萱 杨灿 洪叶挺 SHEN Qian-yi;FAN Jun-jie;HUANG Xin;JIANG Ying;HE Ke-xuan;YANG Can;HONG Ye-ting(School of Laboratory Medicine&Bioengineering,Hangzhou Medical College,Hangzhou 311300;School of Pharmaceutical Sciences,Hangzhou Medical College,Hangzhou 311300;Department of Pathology,Lin’an District First People’s Hospital,Hangzhou 311300)

机构地区：[1]杭州医学院检验医学院生物工程学院,杭州311300 [2]杭州医学院药学院,杭州311300 [3]杭州市临安区第一人民医院病理科,杭州311300

出　　处：《生殖医学杂志》2025年第4期515-524,共10页Journal of Reproductive Medicine

基　　金：国家自然科学基金资助项目(81801513);浙江省高校基本科研经费(KYZD202003)。

摘　　要：目的探究FTO在睾丸组织中的表达定位,并筛选其潜在的调控基因。方法采用qRT-PCR和Western blot技术检测FTO基因在小鼠和大鼠各器官中的表达情况,利用HE染色和免疫组化技术分析FTO在睾丸组织的表达定位;通过慢病毒表达系统构建FTO稳定表达的小鼠精母细胞模型,采用转录组学测序结合qRT-PCR技术筛选FTO潜在的调控基因。结果FTO在睾丸组织中具有较高的mRNA和蛋白表达水平;免疫组化结果进一步显示FTO在睾丸精母细胞中表达量最为显著。基于FTO稳定表达细胞模型转录组学的分析,成功筛选出27个潜在调控基因,并通过qRT-PCR实验验证了6个与精子功能密切相关的基因,分别为Shank3、Dnaaf3、Susd2、Bend5、Ggt5和Svs2。结论FTO在睾丸组织尤其精母细胞中显著表达,提示可能在精子发生过程中扮演关键角色。本研究筛选出的FTO潜在调控基因,为深入理解FTO在精子功能中的作用机制提供了分子依据。Objective:To explore the cellular localization of FTO in testicular tissues and to screen potential regulatory genes using cell models.Methods:The qRT-PCR and western blot analysis were employed to assess the expression profile of FTO gene across various organs in mice and rats.The hematoxylin and eosin(HE)staining and immunohistochemical techniques were utilized to determine the cellular localization of FTO in the testis.A lentiviral system was employed to establish mouse spermatogonial cell models with stable expression of FTO.Both transcriptome sequencing and qRT-PCR were applied to identify genes potentially regulated by FTO.Results:The results demonstrated high levels of mRNA and protein expression of FTO in the testicular tissue,with immunohistochemical analysis indicating particularly elevated expression in spermatogonial cells.Transcriptome analysis of the FTO stably expressing cell models facilitated the identification of 27 candidate regulatory genes.Subsequent qRT-PCR validation confirmed the involvement of 6 genes(Shank3,Dnaaf3,Susd2,Bend5,Ggt5 and Svs2)that were intimately associated with spermatogenic function.Conclusions:FTO was significantly expressed in testicular tissues,especially in spermatogonia,suggesting that it may play a key role in spermatogenesis.In this stady,the potential regulatory genes of FTO,which were screened,provide molecular insights into the mechanisms of FTO in sperm function.

关 键 词：FTO 睾丸 转录组学分析 潜在调控基因 精子功能 

分 类 号：R394.1[医药卫生—医学遗传学]